The proposed Bone Cells and Signaling Core will enable participating investigators to pursue new lines of research in skeletal biology by using state-of-the-art technologies for bone cell isolation and functional assessment, and by advancing their knowledge of how such systems can be applied to problems in the field. The proposed Core will bring together the expertise and acumen of the two Co-Directors Dr. Pajevic Divieti and Dr. Gardella, and powerful new research approaches and assay systems to make for a synergistic resource that will enrich and catalyze the scientific advances of participating investigators. The Core will enable investigators to obtain directly from skeletal tissue, specific types of cells, such as osteoblasts and osteocytes, in purified form, and to propagate these cells for further detailed study. Also enabled will be the means to knock-down specific genes of interest in these cells using shRNA technologies. The Core will further enable analyses of multiple signal transduction and sub-cellular molecular trafficking mechanisms that operate in such cells using sensitive new biochemical assays, high throughput plate reader technologies, and real-time, high-resolution microscopy approaches. As the Core evolves, it will seek to enable and apply new methods, such as CRISPR and TALON technologies to site-specifically alter target genes in cultured cells, scaffolding methods to grow and assay skeletal cells in three dimensions, and multiarray signaling assays to assess novel drug/modulator action, such as biased agonism and signaling selectivity. Each service provided by the Core will be rigorously controlled for quality performance and reliability. The Core will conduct hands-on training sessions by which investigators can be mentored and taught how to perform assays and cell manipulations on their own. It will also establish a seminar series by which new methodologies, approaches, and related scientific progress can be presented and discussed. By providing these services and functions at efficient costs, with high quality, and with rich education, the Core will reduce research barriers for established and junior investigators, and thus help open new lines of investigation and discovery in the skeletal field.